Identification and Characterization of a Large Source of Primary Mesenchymal Stem Cells Tightly Adhered to Bone Surfaces of Human Vertebral Body Marrow Cavities

Abstract

Therapeutic allogeneic mesenchymal stem/stromal cells (MSC) are currently in clinical trials for evaluating their effectiveness in treating many different disease indications. Eventual commercialization for broad distribution will require further improvements in manufacturing processes to economically manufacture MSC at sufficient scales required to satisfy projected demands. A key contributor to the present high cost of goods (COG) for MSC manufacturing is the need to create master cell banks (MCBs) from multiple donors, which leads to variability in large-scale manufacturing runs. Therefore, the availability of large single donor depots of primary MSC would greatly benefit the cell therapy market by reducing costs associated with manufacturing.We have discovered that an abundant population of cells possessing all the hallmarks of MSC is tightly associated with the vertebral body (VB) bone matrix and are only liberated by proteolytic digestion. Here we demonstrate that these vertebral bone-adherent (vBA) MSC possess all the International Society of Cell and Gene Therapy (ISCT)-defined characteristics (e.g., plastic adherence, surface marker expression, and trilineage differentiation) of MSC and, therefore, have termed them vBA-MSC, to distinguish this population from loosely associated MSC recovered through aspiration or rinsing of the bone marrow (BM) compartment.Pilot banking and expansion was performed with vBA-MSC obtained from 3 deceased donors and it was demonstrated that bank sizes averaging 2.9x10 8 +1.35x10 8 vBA-MSC at passage one were obtainable from only 5 g of digested VB bone fragments. Each bank of cells demonstrated robust proliferation through a total of 9 passages without significant reduction in population doubling times. The theoretical total cell yield from the entire amount of bone fragments (approximately 300g) from each donor with limited expansion through 4 passages is 100 trillion (1x10 14 ) vBA-MSC, equating to over 10 5 doses at 10x10 6 cells/kg for an average 70 kg recipient. Thus, we have established a novel and plentiful source of MSC which will benefit the cell therapy market by overcoming manufacturing and regulatory inefficiencies due to donorto-donor variability.

Fabrication of 3D Printed poly(lactic acid) strut and wet-electrospun cellulose nano fiber reinforced chitosan-collagen hydrogel composite scaffolds for meniscus tissue engineering

“…The cells were incubated in chondrogenic, osteogenic, and adipogenic differentiation medium and stained with Alcian blue, Alizarin red, and Oil red O stains, respectively. 53 The undifferentiated rMSCs, incubated in growth medium were observed by light microscopy on 14 days of culture period (Supplemental Figure 1(a)). The cells stained with Alcian blue for chondrogenic differentiation and proteoglycans produced by chondrogenic cells were shown in blue color in the cell cytoplasm (Supplemental Figure 1(b)).…”

“…Certain requirements of a scaffold includes biocompatibility, nontoxicity, good cell-material interactions following physical and chemical properties. 53,54 Biocompatibility is a crucial property which allows optimal cellular interaction, proliferation, and migration for tissue regeneration. The in vitro biocompatibility was assessed on the A3 scaffold which had the best damping capacity, sufficient compression strength and water content in terms of cytotoxicity and its ability to provide desired cellular activity and thus to mimic cartilage tissue.…”

Plasticidade e características gerais das células-tronco mesenquimais derivadas do tecido adiposo humano coletado por lipoaspiração convencional e lipoaspiração por laser

“…A descoberta de novos tecidos fontes para as células-tronco viabilizou a expansão de estudos, principalmente com o objetivo de futuras terapias regenerativa e celular (40).…”

“…O valor de células-tronco obtidas de um grama de tecido adiposo humano pode alcançar até 1x10 6 (39). Outra questão é que as CTMMOs possuem um período de duplicação mais longo, alcançam a senescência mais cedo (9), e constituem apenas 0,01 -0,001% das células nucleadas da medula óssea (18,40).…”